Journal: Kidney Diseases

Article Title: Clinical and Experimental Insights into the Role of NETosis in IgA Nephropathy Pathogenesis

doi: 10.1159/000546343

Figure Lengend Snippet: NETosis promotes TNF-α release from MESs but does not induce cell proliferation. a MES proliferation was not affected by NETosis ( n = 3). b NETosis significantly increased TNF-α secretion ( n = 3). c , d IL-1β and TGF-β levels remained unchanged ( n = 3). Data were analyzed using ANOVA and are presented as mean ± SEM.

Article Snippet: In the experiment evaluating the effects of TNF-α on MESs, MES cells were treated with TNF-α (MCE, HY- P70571 ) at a concentration of 20 ng/mL for 24 h.

Techniques:

Journal: Kidney Diseases

Article Title: Clinical and Experimental Insights into the Role of NETosis in IgA Nephropathy Pathogenesis

doi: 10.1159/000546343

Figure Lengend Snippet: TNF-α stimulation downregulates PPARα in MESs. a Western blot analysis of PPARα, PPARβ/δ, and PPARγ ( n = 3). b–d Relative expression of PPARα, PPARβ/δ, and PPARγ normalized to β-actin. Data are presented as mean ± SEM.

Article Snippet: In the experiment evaluating the effects of TNF-α on MESs, MES cells were treated with TNF-α (MCE, HY- P70571 ) at a concentration of 20 ng/mL for 24 h.

Techniques: Western Blot, Expressing

Journal: Advanced Science

Article Title: Depletion of p75NTR in Schwann Cells Driven by Inflammation Mediates Cutaneous Pain in Psoriasis

doi: 10.1002/advs.202523189

Figure Lengend Snippet: Psoriasis evokes pain‐associated coping behaviors. (A) Representative images of dorsal skin and surrounding hair in Vas‐ and IMQ‐treated mice. Red‐boxed areas are magnified to highlight changes in fur around lesions. (B) Quantification of licking events over 1 h. (C) Representative immunofluorescence images of β3‐tubulin in healthy controls (HC) and psoriasis patients (Ps). (D) The %β3‐tubulin‐positive staining area in C. (E) Representative movement tracks of mice in the open field arena during a 10‐min session. (F) Total distance traveled (m) in the open field. (G) Number of entries into the central zone. (H) Duration spent in the central zone (s). (I) Diagram of the IL‐17A‐induced psoriasis model in C57BL/6 mice injected in hind paws with PBS or IL‐17A (created using Figdraw). (J) H&E staining of PBS‐ and IL‐17A‐injected hind paw skin. (K) Quantification of epidermal thickness in PBS‐ versus IL‐17A‐injected skin. (L) Paw withdrawal threshold (g) measured using Von Frey filaments. (M) Latency to jumping (s) on a hot plate (55°C). (N) Number of hind paw lifts during 5‐min cold plate exposure (4°C). (O) Total duration of hind paw lifting (s) during cold plate exposure. Scale bars as indicated. ns, not significant; * p < 0.05, ** p < 0.01, *** p < 0.001, and **** p < 0.0001; according to unpaired Student's t ‐test (B, D, F, G, H, K, L, M, N, O). n = 3 (D), n = 6 (B, F, G, H, K, L, M, N, O).

Article Snippet: Human IL‐17A protein (HY‐ P70527 , MCE), Rat IL‐17A protein (HY‐ P78556 , MCE), human β‐NGF (HZ‐1222, Proteintech), Tanezumab (HY‐P99221, MCE), ML385 (HY‐100523, MCE) and CPZ (HY‐12708, MCE) were used for cell stimulation as specified in the experiments. siRNAs targeting human p75NTR ( NGFR ) and a negative control siNC were synthesized commercially.

Techniques: Immunofluorescence, Staining, Injection

Journal: Advanced Science

Article Title: Depletion of p75NTR in Schwann Cells Driven by Inflammation Mediates Cutaneous Pain in Psoriasis

doi: 10.1002/advs.202523189

Figure Lengend Snippet: Psoriatic lesions exhibit elevated NGF with cutaneous TrkA loss and DRG accumulation. (A) Representative fluorescent immunohistochemistry images and quantification of NGF mean fluorescence intensity of NGF in healthy controls (HC) and psoriasis patients (Ps). (B) Quantification of NGF levels in skin tissues from healthy controls (HC) and psoriasis patients. (C) Western blot analysis of NGF and IL‐17A expression in dorsal skin of Vas‐ and IMQ‐treated mice. (D) Statistical analysis of NGF/IL‐17A expression from C. (E) Western blot analysis of p‐TrkA and TrkA protein expression in dorsal skin of Vas‐ and IMQ‐treated mice. (F) Quantification of p‐TrkA and TrkA expression from E. (G) Schematic of neurons with cell bodies in the dorsal root ganglia (DRG) projecting axons to the skin. (H) Western blot analysis of protein expression in DRG of Vas‐ and IMQ‐treated mice. (I) Quantification of protein expression from H. (J) Relative c‐Fos mRNA expression in DRG from Vas‐ and IMQ‐treated mice. In (E), the two arrow‐indicated bands together represent the total TrkA signal in mouse skin and were quantified in (F). Scale bar as indicated. * p < 0.05, ** p < 0.01, *** p < 0.001, and **** p < 0.0001; unpaired Student's t ‐test (A, B, D, F, I, J); n = 3 (A, B, I, J), n = 6 (D, F).

Article Snippet: Human IL‐17A protein (HY‐ P70527 , MCE), Rat IL‐17A protein (HY‐ P78556 , MCE), human β‐NGF (HZ‐1222, Proteintech), Tanezumab (HY‐P99221, MCE), ML385 (HY‐100523, MCE) and CPZ (HY‐12708, MCE) were used for cell stimulation as specified in the experiments. siRNAs targeting human p75NTR ( NGFR ) and a negative control siNC were synthesized commercially.

Techniques: Immunohistochemistry, Fluorescence, Western Blot, Expressing

Journal: Advanced Science

Article Title: Depletion of p75NTR in Schwann Cells Driven by Inflammation Mediates Cutaneous Pain in Psoriasis

doi: 10.1002/advs.202523189

Figure Lengend Snippet: IL‐17A and NGF are associated with MLC2‐Linked cytoskeletal remodeling in Schwann cells. (A) Representative TEM images of myelinated bundles in the dorsal skin of Vas‐ and IMQ‐treated mice. (B) TEM images of myelinated bundles in PBS‐ or IL‐17A‐injected hind paws. (C) TEM images of glio‐neural complexes in PBS‐ or IL‐17A‐injected hind paws. (D) TEM images of glio‐neural complexes in healthy controls (HC) and psoriasis patients (Ps). Red arrows indicate Schwann cell nuclei in (A, B). Glio‐neural complexes at the dermal–epidermal junction were pseudo‐colored: axons (green), Schwann cells and processes (red) in (C, D). (E) Representative fluorescence images of Schwann cells under PBS (control) or IL‐17A/NGF treatment, stained with phalloidin. (F–H) Quantification of Schwann cell morphology index (F), relative cell area (G), and F‐actin mean intensity (H) under control conditions. (I, J) Schwann cells were stimulated with NGF (100 ng/mL, I) or IL‐17A (20 ng/mL, J) at the indicated time points, and total MLC2 and p‐MLC2 levels were analyzed by Western blot. Scale bars as indicated. ns, not significant; * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001; one‐way ANOVA with post hoc Tukey (F–H), n = 5 (F–H), n = 3 (I, J).

Article Snippet: Human IL‐17A protein (HY‐ P70527 , MCE), Rat IL‐17A protein (HY‐ P78556 , MCE), human β‐NGF (HZ‐1222, Proteintech), Tanezumab (HY‐P99221, MCE), ML385 (HY‐100523, MCE) and CPZ (HY‐12708, MCE) were used for cell stimulation as specified in the experiments. siRNAs targeting human p75NTR ( NGFR ) and a negative control siNC were synthesized commercially.

Techniques: Injection, Fluorescence, Control, Staining, Western Blot

Journal: Advanced Science

Article Title: Depletion of p75NTR in Schwann Cells Driven by Inflammation Mediates Cutaneous Pain in Psoriasis

doi: 10.1002/advs.202523189

Figure Lengend Snippet: Schwann cell proliferation in psoriatic‐like lesions. (A) Representative immunofluorescence images of mouse skin stained for SOX10 (Schwann cells, red), β3‐Tubulin (nerve fibers, green), and DAPI (nuclei, blue). (B) Western blot analysis of Schwann cell‐ and nerve‐related proteins in Vas‐ and IMQ‐treated mice. (C) Quantification of S100B, SOX10, MBP, and β3‐Tubulin from B. (D) Workflow schematic for isolation of primary Schwann cells from skin. (E) Representative flow cytometry plot showing identification of Schwann cells gated as SOX10 + S100B + . (F) Quantification of SOX10 + S100B + cells as a percentage of live singlets. (G) Representative immunofluorescence images of Ki67+ RSC96 cells (green) after IL‐17A stimulation (20 ng/mL); nuclei labeled with DAPI (blue). (H) Quantification of Ki67+ RSC96 cells. Scale bars as indicated. * p < 0.05, *** p < 0.001, **** p < 0.0001; unpaired Student's t ‐test (C, F, H); n = 6 (C), n = 3 (F, H).

Article Snippet: Human IL‐17A protein (HY‐ P70527 , MCE), Rat IL‐17A protein (HY‐ P78556 , MCE), human β‐NGF (HZ‐1222, Proteintech), Tanezumab (HY‐P99221, MCE), ML385 (HY‐100523, MCE) and CPZ (HY‐12708, MCE) were used for cell stimulation as specified in the experiments. siRNAs targeting human p75NTR ( NGFR ) and a negative control siNC were synthesized commercially.

Techniques: Immunofluorescence, Staining, Western Blot, Isolation, Flow Cytometry, Labeling

Journal: Advanced Science

Article Title: Depletion of p75NTR in Schwann Cells Driven by Inflammation Mediates Cutaneous Pain in Psoriasis

doi: 10.1002/advs.202523189

Figure Lengend Snippet: Schwann cells attenuate NGF‐Driven DRG TrkA signaling via p75NTR, while IL‐17 potentiates the response. (A) PCR and agarose gel electrophoresis analysis of p75NTR and TrkA in Schwann cells (SCs). (B, C) Validation of p75NTR knockdown in primary skin SCs and SCs using siRNA. (D, E) Western blot analysis of p75NTR expression in primary skin SCs and SCs treated with increasing NGF concentrations; accompanying quantification is shown. (F) Western blot and quantification of p75NTR in SCs treated with CHX (70 µg/mL) for the indicated times, with or without NGF (12 h). (G) Schematic of the experimental design for SCs, PC12 cells, and co‐culture under ±NGF and ±IL‐17A conditions (created using Figdraw). (H) Western blot of p‐TrkA and total TrkA in indicated cells after 5‐min NGF (100 ng/mL) stimulation; quantification of p‐TrkA/TrkA ratio shows reduced TrkA phosphorylation in the co‐culture relative to PC12 cells alone. (I) Immunofluorescence of p‐TrkA (red), GFP (green; lentiviral marker in PC12), and DAPI (blue); arrowheads indicate GFP‐negative/DAPI‐positive nuclei corresponding to SCs in the co‐culture. (J) Western blot and quantification of TrkA phosphorylation in co‐cultures treated with NGF and IL‐17A. (K) Schematic of DRG neuron and RSC96 co‐culture assay ±IL‐17A; DRG neurons cultured independently served as controls (created using Figdraw). (L) Representative immunofluorescence images of neurite outgrowth in co‐culture. (M) Quantification of average neurite area per cell. Scale bars as indicated. ns, not significant; * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001; one‐way ANOVA with post hoc Tukey (D, E), two‐way ANOVA with post hoc Tukey (F, M), unpaired Student's t ‐tests (H, J), n = 3 (D, E, F, H, J), n = 17 (M).

Article Snippet: Human IL‐17A protein (HY‐ P70527 , MCE), Rat IL‐17A protein (HY‐ P78556 , MCE), human β‐NGF (HZ‐1222, Proteintech), Tanezumab (HY‐P99221, MCE), ML385 (HY‐100523, MCE) and CPZ (HY‐12708, MCE) were used for cell stimulation as specified in the experiments. siRNAs targeting human p75NTR ( NGFR ) and a negative control siNC were synthesized commercially.

Techniques: Agarose Gel Electrophoresis, Biomarker Discovery, Knockdown, Western Blot, Expressing, Co-Culture Assay, Phospho-proteomics, Immunofluorescence, Marker, Co-culture Assay, Cell Culture

Journal: Advanced Science

Article Title: Depletion of p75NTR in Schwann Cells Driven by Inflammation Mediates Cutaneous Pain in Psoriasis

doi: 10.1002/advs.202523189

Figure Lengend Snippet: IL‐17A enhances NGF secretion and p75NTR regulation in Schwann cells. (A, B) Western blot and quantification of p75NTR and NGF in SCs stimulated with IL‐17A at the indicated time points. (C, D) Dot plots showing the expression of p75NTR and Ngf in SCs from the Vas and IMQ groups. Dot size indicates the percentage of cells expressing each gene, and dot color represents the average expression level. (E) Changes in p75NTR protein levels in SCs treated with IL‐17A (20 ng/mL) in the presence of bafilomycin A1 (Baf A1, 40 nM) or MG132 (20 µM). (F) Western blot and quantification of NGF expression in RSC96 cells treated with IL‐17A for the indicated times. (G) Western blot of NGF in supernatants of RSC96 cells stimulated with IL‐17A (20 ng/mL, 12 h); GM130 served as a negative control to confirm the absence of cellular contamination. (H) Schematic of PC12 treatment with RSC96‐derived conditioned media stimulated by IL‐17A (created using Figdraw). (I) Western blot of PC12 lysates after 5‐min exposure to RSC96‐conditioned media. (J) Quantification of protein levels in I. ns, not significant; * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001; one‐way ANOVA with post hoc Tukey (B, E, F, J), unpaired Student's t ‐test (G), n = 3.

Article Snippet: Human IL‐17A protein (HY‐ P70527 , MCE), Rat IL‐17A protein (HY‐ P78556 , MCE), human β‐NGF (HZ‐1222, Proteintech), Tanezumab (HY‐P99221, MCE), ML385 (HY‐100523, MCE) and CPZ (HY‐12708, MCE) were used for cell stimulation as specified in the experiments. siRNAs targeting human p75NTR ( NGFR ) and a negative control siNC were synthesized commercially.

Techniques: Western Blot, Expressing, Negative Control, Derivative Assay

Journal: Advanced Science

Article Title: Depletion of p75NTR in Schwann Cells Driven by Inflammation Mediates Cutaneous Pain in Psoriasis

doi: 10.1002/advs.202523189

Figure Lengend Snippet: Keap1 downregulation mediates IL‐17A‐Induced Nrf2 activation and NGF upregulation. (A) mRNA expression of Nrf2 and Keap1 in RSC96 cells treated with IL‐17A (20 ng/mL) at indicated times. (B) Western blot analysis of Nrf2 and Keap1 protein levels in RSC96 cells under IL‐17A (20 ng/mL) treatment. (C) Quantification of protein expression from B. (D) Representative immunofluorescence images of Nrf2 (green) in RSC96 cells. (E) Western blot of nuclear and cytosolic Nrf2 in RSC96; Lamin B and GAPDH served as nuclear and cytoplasmic loading controls. (F) Quantification of nuclear Nrf2 from E. (G) Western blot of RSC96 lysates treated with IL‐17A and ML385 (5 µM). (H) Quantification of protein levels in G. ns, not significant; * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001; unpaired Student's t ‐test (F), one‐way ANOVA with post hoc Tukey (A, C, H), n = 3.

Article Snippet: Human IL‐17A protein (HY‐ P70527 , MCE), Rat IL‐17A protein (HY‐ P78556 , MCE), human β‐NGF (HZ‐1222, Proteintech), Tanezumab (HY‐P99221, MCE), ML385 (HY‐100523, MCE) and CPZ (HY‐12708, MCE) were used for cell stimulation as specified in the experiments. siRNAs targeting human p75NTR ( NGFR ) and a negative control siNC were synthesized commercially.

Techniques: Activation Assay, Expressing, Western Blot, Immunofluorescence

Journal: Advanced Science

Article Title: Depletion of p75NTR in Schwann Cells Driven by Inflammation Mediates Cutaneous Pain in Psoriasis

doi: 10.1002/advs.202523189

Figure Lengend Snippet: Schwann cell–targeted p75NTR overexpression attenuates IL‐17A–evoked nociceptive sensitization. (A) Schematic of the AAV expression cassette. (B) Experimental timeline: plantar AAV injection on day 1; in vivo imaging validation on day 21; repeated plantar IL‐17A (or PBS) injections on days 21, 23, 25, and 27; followed by behavioral testing. (C) Representative in vivo fluorescence imaging showing localized reporter signals at the hind‐paw injection sites in AAV‐Vector and AAV‐p75NTR mice. (D) Immunoblotting of p75NTR in mouse plantar skin lysates with GAPDH as a loading control, and quantification of relative p75NTR protein levels. (E) Mechanical sensitivity assessed by von Frey test (withdrawal threshold, g) in four groups. (F,G) Cold plate test showing the number of paw‐lifting events (F) and lifting duration (G). (H) Hot plate test showing thermal latency. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001; unpaired Student's t ‐test (D), one‐way ANOVA with post hoc Tukey (E–H), n = 4 (D), n = 8 (E‐H).

Article Snippet: Human IL‐17A protein (HY‐ P70527 , MCE), Rat IL‐17A protein (HY‐ P78556 , MCE), human β‐NGF (HZ‐1222, Proteintech), Tanezumab (HY‐P99221, MCE), ML385 (HY‐100523, MCE) and CPZ (HY‐12708, MCE) were used for cell stimulation as specified in the experiments. siRNAs targeting human p75NTR ( NGFR ) and a negative control siNC were synthesized commercially.

Techniques: Over Expression, Expressing, Injection, In Vivo Imaging, Biomarker Discovery, In Vivo, Fluorescence, Imaging, Plasmid Preparation, Western Blot, Control, Hot Plate Test

Journal: Journal of Nanobiotechnology

Article Title: Inverse-designed plasmonic biosensors with LSPR-SPP-wood anomaly coupling enhanced for biomolecular analysis

doi: 10.1186/s12951-025-03930-w

Figure Lengend Snippet: High-throughput affinity sensing platform for ADC drug development. ( a ) The association process of incubating PD-1 on the chip to detect Sintilimab. ( b ) The long-time dissociation process of PD-1 with Sintilimab. ( c ) Dynamic real-time and fitting curves of the association and dissociation phases of the interaction between PD-1 and Sintilimab. ( d ) Schematic diagram of the high-throughput antibody affinity detection protocol, with each group, repeated three times in the three samples tested, and the concentration of A-H in each column decreasing incrementally. ( e ) Schematic diagram of the main events in the antibody drug fishing and affinity detection process. ( f-h ) Dynamic real-time and fitting curves of the association and dissociation phases for Polatuzumab with CD79B, Adalimumab with TNF-α, and Sacituzumab govitecan with TROP2, detected using the unoptimized sensor. ( i-k ) Dynamic real-time and fitting curves of the association and dissociation phases for Polatuzumab with CD79B, Adalimumab with TNF-α, and Sacituzumab govitecan with TROP2, detected using the optimized IDMM-SPR sensor

Article Snippet: Bovine serum albumin (BSA), Phosphate-buffered saline (PBST), 11-mercaptoundecanoic acid (MUA), 1-ethyl3-(dimethylaminopropyl) carbodiimide (EDC), N-hydroxysuccinimide (NHS), Dimethyl Sulfoxide (DMSO), Fish gelatin,, ethanolamine, and phosphate-buffered saline (PBS) buffer, Polyethylene glycol 2000 (PEG) were purchased from Sigma-Aldrich; Protein A, IgG, CRP, and CRP ab, hTGFBR1 were purchased from monoclonal anti-CRP capture and detection antibodies were purchased from Beijing Sino Biological, Inc. (Beijing, China); Sintilimab, Adalimumab, Polatuzumab, Sacituzumab, PD-1, TNF-α, CD79B, TROP2, Quercetin were purchased from MedChemExpress.

Techniques: High Throughput Screening Assay, Concentration Assay

Journal: Theranostics

Article Title: CEBPB-high dormant tumor cells drive immune evasion via S100A8 orchestrated tumor-associated macrophages reprogramming

doi: 10.7150/thno.124789

Figure Lengend Snippet: CEBPB transcriptionally activates S100A8 to promote M2 macrophage polarization. A , Venn diagram of three datasets identifying four common secretory factors: differentially expressed genes from CEBPB-overexpressing 4T07 cells bulk RNA-seq, CEBPB binding peaks in 4T07 cells from anti-CEBPB ChIP-seq, and the profile of mouse secretory proteins. B , CEBPB binding peaks for S100A8 and Cxcl1 from anti-CEBPB ChIP-seq in 4T07 cells. C , ChIP-qPCR showing the CEBPB binding regions in the S100a8 promoter or upstream regions of Cxcl1 of 4T07 and EMT6 cells. D , Relative luciferase activity in CEBPB-overexpressing HEK293T cells with the S100a8 promoter WT or MUT, respectively. E-F , mRNA expression of S100a8 or Cxcl1 in 4T07 cells with CEBPB overexpression (E) or knockdown (F). G , Protein expression of S100A8 in 4T07 cells with or without CEBPB overexpression. H , Representative multiplex immunohistochemistry images of Ki67, CEBPB, and S100A8 staining in a TMA of TNBC patient tumor smaples ( n = 80). I , mRNA expression of S100a8 in eGFP -/lo and eGFP +/hi tumor cells sorted from 4T07-TetOn or EMT6-TetOn tumor masses. J-K , Percentages of CD206 (J) and MHCII (K) expressing BMDMs with the addition of rS100A8 (100 or 500 ng/mL) or rCXCL1 (100 or 500 ng/mL) for 48 hours. L-N , Tumor volume (L), image (M) and weight (N) of CEBPB-overexpressing with or without S100A8 knockdown EMT6 allografts in BALB/c mice ( n = 5). O-R , Flow cytometry analysis of CD8 + T cells (O), CD11b + F4/80 + cells (P), CD206 + macrophages (Q), and MHCII + macrophages (R) infiltrated in tumors developed in (L). n.s., not significant, *, P < 0.05, **, P < 0.01, ***, P < 0.001.

Article Snippet: BMDMs or PBMC-M φ cells were co-cultured in conditional medium supplemented with 10% FBS and 1% penicillin-streptomycin, or in complete DMEM or 1640 medium supplemented with rS100A8 (MCE, #HY- P73671 for BMDMs and #HY- P70531 for PBMC-M φ ) or rCXCL1 (MCE, #HY-P7188 for BMDMs) for 48 hours.

Techniques: RNA Sequencing, Binding Assay, ChIP-sequencing, ChIP-qPCR, Luciferase, Activity Assay, Expressing, Over Expression, Knockdown, Multiplex Assay, Immunohistochemistry, Staining, Flow Cytometry

Journal: Theranostics

Article Title: CEBPB-high dormant tumor cells drive immune evasion via S100A8 orchestrated tumor-associated macrophages reprogramming

doi: 10.7150/thno.124789

Figure Lengend Snippet: The S100A8 inhibitor remodels TME and improves ICB efficacy. A-B , Percentages of CD206 expressing in BMDMs (A) or PBMC-Mφ (B) with the addition of paquinimod (PAQ, 5 µg/mL) and/or rS100A8 (500 ng/mL) for 48 hours. C , Migration assay of BMDMs in DMEM with the addition of PAQ (5 µg/mL) and/or rS100A8 (500 ng/mL) for 24 hours. D-E , Percentages of CD206 expressing in BMDMs (D) or PBMC-Mφ (E) co-cultured with CM and/or PAQ (5 µg/mL) for 48 hours, including CM from 4T07 (D) or MDA-MB-231 (E) with or without CEBPB overexpression. F-H , Tumor volume (F), image (G) and weight (H) of EMT6 allografts in BALB/c mice ( n = 5). Tumor-bearing mice were randomly divided into four groups until tumor volume reached approximately 100 mm 3 . I-L , Flow cytometry analysis of CD8 + T cells (I), CD11b + F4/80 + macrophages (J), CD206 + macrophages (K) MHCII + macrophages (L) infiltrated in tumors developed of the indicated groups. M , Kaplan-Meier analysis of S100A8 expression in a public cohort with 238 patients receiving anti-PD-1 therapy. n.s., not significant, *, P < 0.05, **, P < 0.01, ***, P < 0.001 .

Article Snippet: BMDMs or PBMC-M φ cells were co-cultured in conditional medium supplemented with 10% FBS and 1% penicillin-streptomycin, or in complete DMEM or 1640 medium supplemented with rS100A8 (MCE, #HY- P73671 for BMDMs and #HY- P70531 for PBMC-M φ ) or rCXCL1 (MCE, #HY-P7188 for BMDMs) for 48 hours.

Techniques: Expressing, Migration, Cell Culture, Over Expression, Flow Cytometry

Journal: Cancer Medicine

Article Title: The MIR181A2HG / miR ‐5680/ VCAN ‐ CD44 Axis Regulates Gastric Cancer Lymph Node Metastasis by Promoting M2 Macrophage Polarization

doi: 10.1002/cam4.70600

Figure Lengend Snippet: GC cell CM induces M2‐like polarization of macrophages, promoting lymphangiogenesis. (A) Immunofluorescence shows changes in the M2‐type macrophage marker (CD163) after treatment with conditioned medium from different treatment groups: Si‐NC (control group) and si‐#1 (si‐MIR181A2HG‐1) (scale bar, 50 μm). (B) Quantification of the Proportion of M2 Macrophages (CD163) Among Total Macrophages (CD68). (C) Schematic of the co‐culture model of GC cells and macrophages promoting HLECs lymphangiogenesis. (D) Typical images of macrophage morphological changes after treatment with PMA and GC cell CM (scale bar, 20 μm). (E–H) qRT‐PCR detection of typical M2 markers (CD163, CD206, and IL‐10) and M1 markers (iNOS, IL‐6, and TNFα) in PMA‐treated THP‐1 cells cultured with CM from different treatment groups (si‐NC, si‐#1 and si‐#2) of GC cells. (I) ELISA experiment detects the content of VEGF‐C in the CM after culturing macrophages with CM from different treatment groups (si‐NC, si‐#1 and si‐#2) of GC cells. (J) Immunofluorescence shows changes in the macrophage marker (CD163) after treatment with CM from different treatment groups: Si‐NC, si‐#1 and si‐#2 (scale bar, 50 μm). (K) Tube formation and transwell experiments detect the effects of tube formation and migration invasion ability of HLECS by macrophage CM from different treatment groups: Si‐NC, si‐#1 and si‐#1 + recombinant VEGF‐C protein. Statistical significance was assessed using two‐tailed t ‐tests. * p < 0.05, ** p < 0.01, *** p < 0.001 (scale bar, 50 μm).

Article Snippet: On the other hand, HLECs are cultured in Endothelial Cell Medium (ECM) supplemented with 10% serum and 0.1% endothelial cell growth factor. recombinant VEGF‐C and VCAN protein were purchased from MedChemExpress (New Jersey, USA), Calcein AM were purchased from Beyotime (Shanghai, China), and Phorbol‐12‐myristate‐13‐acetate (PMA) was sourced from SIGMA (Missouri, USA).

Techniques: Immunofluorescence, Marker, Control, Co-Culture Assay, Quantitative RT-PCR, Cell Culture, Enzyme-linked Immunosorbent Assay, Migration, Recombinant, Two Tailed Test

Journal: Cancer Medicine

Article Title: The MIR181A2HG / miR ‐5680/ VCAN ‐ CD44 Axis Regulates Gastric Cancer Lymph Node Metastasis by Promoting M2 Macrophage Polarization

doi: 10.1002/cam4.70600

Figure Lengend Snippet: The MIR181A2HG/miR‐5680/VCAN axis affects lymphangiogenesis by influencing the polarization of M2‐type macrophages. (A) Elisa detection of VCAN protein expression levels in different treatment groups of GC cells: NC, si‐#1 (si‐MIR181A2HG‐1), si‐#1 + miR‐5680 inhibitor, si‐#1 + miR‐5680 inhibitor +si‐VCAN. (B–E) qRT‐PCR detection of typical M2 markers (CD163, CD206, and IL‐10) and M1 markers (iNOS, IL‐6, and TNFα) in PMA‐treated THP‐1 cells cultured with CM from different treatment groups (NC, si‐#1 (si‐MIR181A2HG‐1), si‐#1 + miR‐5680 inhibitor, si‐#1 + miR‐5680 inhibitor +si‐VCAN) of GC cells. (F) ELISA experiment detects the content of VEGF‐C in the supernatant after culturing macrophages with CM from different treatment groups (NC, si‐#1 (si‐MIR181A2HG‐1), si‐#1 + miR‐5680 inhibitor, si‐#1 + miR‐5680 inhibitor +si‐VCAN) of GC cells. (G–K) Tube formation and transwell experiments detect the effects and quantification of tube formation and migration invasion ability of HLECS by macrophage CM from different treatment groups: NC, si‐#1 (si‐MIR181A2HG‐1), si‐#1 + miR‐5680 inhibitor, si‐#1 + miR‐5680 inhibitor +si‐VCAN. Statistical significance was assessed using two‐tailed t ‐tests. * p < 0.05, ** p < 0.01, *** p < 0.001 (scale bar, 50 μm).

Article Snippet: On the other hand, HLECs are cultured in Endothelial Cell Medium (ECM) supplemented with 10% serum and 0.1% endothelial cell growth factor. recombinant VEGF‐C and VCAN protein were purchased from MedChemExpress (New Jersey, USA), Calcein AM were purchased from Beyotime (Shanghai, China), and Phorbol‐12‐myristate‐13‐acetate (PMA) was sourced from SIGMA (Missouri, USA).

Techniques: Enzyme-linked Immunosorbent Assay, Expressing, Quantitative RT-PCR, Cell Culture, Migration, Two Tailed Test

Journal: Cancer Medicine

Article Title: The MIR181A2HG / miR ‐5680/ VCAN ‐ CD44 Axis Regulates Gastric Cancer Lymph Node Metastasis by Promoting M2 Macrophage Polarization

doi: 10.1002/cam4.70600

Figure Lengend Snippet: VCAN Induces M2 Macrophage Activation via Binding to CD44. (A) Prediction of VCAN interacting proteins based on online databases String, GeneMANIA, Hitpredict, Biogrid, Hint. (B) String database predicts CD44 interacts with VCAN. Purple line: Experimentally determined. Green line: Text mining. Black line: Co‐expression. (C) Co‐IP experiment detects the mutual binding of CD44 and VCAN in polarized macrophages. (D) Immunofluorescence detects the co‐localization of CD44 and VCAN in polarized macrophages (scale bar, 50 μm). (E–H) qRT‐PCR detection of typical M2 markers (CD163, CD206, and IL‐10) and M1 markers (iNOS, IL‐6, and TNFα) in PMA‐treated THP‐1 cells cultured with CM from different treatment groups (NC, VCAN recombinant protein, VCAN recombinant protein + anti‐CD44) of GC cells. (I) Immunofluorescence shows changes in the macrophage marker (CD163) after treatment with CM from different treatment groups: NC, VCAN recombinant protein, VCAN recombinant protein + anti‐CD44 (scale bar, 50 μm). (J) ELISA experiment detects the content of VEGF‐C in the supernatant after culturing macrophages with CM from different treatment groups (NC, VCAN recombinant protein, VCAN recombinant protein + anti‐CD44) of GC cells. (K) Tube formation and transwell experiments detect the effects of macrophage CM from different treatment groups (NC, VCAN recombinant protein, VCAN recombinant protein + anti‐CD44) on the tube formation and migration invasion ability of HLECS (scale bar, 50 μm). (L) Co‐IP experiment detects the mutual binding of CD44 and VCAN in GC cells. * p < 0.05, ** p < 0.01, *** p < 0.001.

Article Snippet: On the other hand, HLECs are cultured in Endothelial Cell Medium (ECM) supplemented with 10% serum and 0.1% endothelial cell growth factor. recombinant VEGF‐C and VCAN protein were purchased from MedChemExpress (New Jersey, USA), Calcein AM were purchased from Beyotime (Shanghai, China), and Phorbol‐12‐myristate‐13‐acetate (PMA) was sourced from SIGMA (Missouri, USA).

Techniques: Activation Assay, Binding Assay, Expressing, Co-Immunoprecipitation Assay, Immunofluorescence, Quantitative RT-PCR, Cell Culture, Recombinant, Marker, Enzyme-linked Immunosorbent Assay, Migration

Journal: Cell Discovery

Article Title: Integrated transcriptome profiling of plasma exosomes reveals molecular stratification of exocrine and endocrine disorders and S100A8-mediated cell interactions in chronic pancreatitis

doi: 10.1038/s41421-025-00832-x

Figure Lengend Snippet: a Overview of the filtering strategy. b The screened miRNA (mir-24-3p)–mRNA interaction network. c The expression of miR-24-3p in each COCA subtype and normal samples. d S100A8 expression in each COCA subtype and normal samples. e Schematic diagram of the CP mouse model construction, single-cell data processing, and estimation of exosome secretion activity using the sEVtrans algorithm. f Left: UMAP plot of all cell types and identified exosomes (sEVs) in the single-cell sequencing data. Right: proportion of exosomes from the CP group and NC group. g UMAP plot showing the exosome secretion activity index (ESAI) value distributions among different cells. h Violin plot showing the ESAI distributions in different cell types. **** P < 0.0001. i Violin plot showing the expression of S100A8 in different cell types. j Representative images of Ly6G, F4/80 and S100A8 immunofluorescence in the mouse CP model. k Schematic diagram of the CP mouse model constructed with intraperitoneal injection of a neutrophil depletion antibody (anti-Ly6g) compared with that generated by isotype injection. l Representative flow cytometry image after establishment of the CP model compared with that after mock establishment via isotype injection. m Neutrophil counts in CD45 + blood cells in CP mice after model establishment compared with mock establishment. n mRNA expression of exosomal S100A8 in the CP mice after model establishment compared with mock establishment.

Article Snippet: Different level of S100A8 protein (HY- P70531 , MCE company, USA) was used to stimulate the HPSC cells.

Techniques: Expressing, Activity Assay, Sequencing, Immunofluorescence, Construct, Injection, Generated, Flow Cytometry

Journal: Cell Discovery

Article Title: Integrated transcriptome profiling of plasma exosomes reveals molecular stratification of exocrine and endocrine disorders and S100A8-mediated cell interactions in chronic pancreatitis

doi: 10.1038/s41421-025-00832-x

Figure Lengend Snippet: a Exosomes loaded with PKH26 dye can be taken up by HL-60 neutrophils, including the exosome-supplemented group and the empty control group. b S100A8 mRNA expression in HL-60 cells after transfection with miR-24-3p or S100A8. n = 3 per group. c Protein expression of S100A8 in HL-60 cells after transfection with miR-24-3p mimic or inhibitor. n = 3 per group. d Design of the luciferase reporter vector with sequencing alignment of miR-24-3p with the 3’-UTR of the S100A8 gene. e Luciferase activity of the human S100A8 3’-UTR in HEK293T cells. Cells were transfected with empty plasmid (NC), WT, or mutant 3’-UTR (Mut) plasmids together with miR-24-3p mimics or inhibitors. n = 3 per group. f Schematic diagram of the neutrophil and pancreatic stellate cell co-culture system. Relative protein levels of collagen-1 in pancreatic stellar cells after co-culture with mir-24-3p mimic ( g ), inhibitor ( h ), or paquinimod ( i ). n = 3 per group. j The protein levels of collagen-1 and FN in pancreatic stellate cells after exogenous supplementation with S100A8. n = 3 per group. k Schematic diagram of paquinimod administration in the CP mouse model. l Representative HE and Masson sections from CP model mice with or without paquinimod injection (left) and the pathological scores of the two groups. n = 5 per group. m The protein levels of collagen-1 and FN in pancreatic stellate cells after exogenous supplementation with S100A8 (1 µg/mL) and transfection with CD36, TLR4, RAGE or NC siRNA. n = 3 per group. n Representative fluorescence images of ROS in pancreatic stellate cells from the control group, S100A8 (1 µg/mL) group, S100A8 (1 µg/mL) + 5 mM NAC group, and S100A8 (1 µg/mL) + 10 mM NAC group. o The protein levels of collagen-1 and FN in pancreatic stellate cells after exogenous supplementation with S100A8 (1 µg/mL) or NAC (5 mM or 10 mM). n = 3 per group.

Article Snippet: Different level of S100A8 protein (HY- P70531 , MCE company, USA) was used to stimulate the HPSC cells.

Techniques: Control, Expressing, Transfection, Luciferase, Plasmid Preparation, Sequencing, Activity Assay, Mutagenesis, Co-Culture Assay, Injection, Fluorescence

Journal: Cell Discovery

Article Title: Integrated transcriptome profiling of plasma exosomes reveals molecular stratification of exocrine and endocrine disorders and S100A8-mediated cell interactions in chronic pancreatitis

doi: 10.1038/s41421-025-00832-x

Figure Lengend Snippet: a mRNA expression of IL-1B, IL6, and TNF in RAW264.7 cells after exogenous supplementation with S100A8 (1 µg/mL) and transfection with or without TLR4 siRNA. n = 3 per group. b Schematic diagram of the macrophage and pancreatic stellate cell co-culture system. c Relative protein levels of BCL-2 and Bax in MIN6 cells after culture with RAW264.7 cells, TLR4-knockdown RAW264.7 cells, and/or exogenous supplementation with S100A8 (1 µg/mL). n = 3 per group. d Representative immunofluorescence image and TUNEL-positive cell counts of MIN6 cells after co-culture with RAW264.7 cells, TLR4-knockdown RAW264.7 cells, and/or exogenous supplementation with S100A8 (1 µg/mL). n = 5 per group.

Article Snippet: Different level of S100A8 protein (HY- P70531 , MCE company, USA) was used to stimulate the HPSC cells.

Techniques: Expressing, Transfection, Co-Culture Assay, Knockdown, Immunofluorescence, TUNEL Assay